Vascular endothelial growth factor mRNA in pericytes is upregulated by phorbol myristate acetate.
Increased microvascular permeability, which occurs in conditions such as the adult respiratory distress syndrome and diabetes mellitus, is related to physicochemical alterations in the microvascular barrier. We postulate that, in part, capillary pericytes affect microvascular permeability via production of a vasoactive cytokine, viz, vascular endothelial growth factor (VEGF), also known as vascular permeability factor. The goal of the present study was to evaluate the effects of phorbol myristate acetate (PMA), a substance known to produce nonhydrostatic pulmonary edema in intact animals, on VEGF gene expression in pericyte cultures. Microvascular pericytes were isolated from bovine retinas using magnetic microspheres coated with 3G5 monoclonal antibody. Pericyte identity was confirmed both morphologically and by immunostaining for alpha-smooth muscle actin and 3G5 ganglioside. The cultured pericytes were stimulated with N(omega)-nitro-L-arginine methyl ester (L-NAME, 1 x 10(-4) mmol/L), angiotensin II (1 x 10(-6) mmol/L), and PMA (5 x 10(-8) mmol/L), selected because of their ability to upregulate VEGF mRNA expressions in other cell types. Northern blot analysis was performed using [32P]dCTP labeled human VEGF cDNA (Genentech). Lane-loading differences were normalized using mouse GAPDH control cDNA probe. VEGF mRNA expression was upregulated by PMA (10(-9) to 10(-6) mol/L) in a dose-dependent manner, whereas neither L-NAME nor angiotensin II affected VEGF mRNA expression in pericytes. These results support the hypothesis that pericytes increase permeability of the endothelial barrier through increased VEGF production.
Kim
Y
Y
Department of Internal Medicine, Saint Louis University School of Medicine, Mo 63104, USA.
Imdad
R Y
RY
Stephenson
A H
AH
Sprague
R S
RS
Lonigro
A J
AJ
D000804:Angiotensin II / Q000494:pharmacology; D000818:Animals; D000911:Antibodies, Monoclonal; D001160:Arterioles / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D002417:Cattle; D016228:Endothelial Growth Factors / Q000096:biosynthesis*; D006801:Humans; D018189:Immunomagnetic Separation; D008222:Lymphokines / Q000096:biosynthesis*; D051379:Mice; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D012333:RNA, Messenger / Q000096:biosynthesis; D012171:Retinal Vessels / Q000166:cytology / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D014158:Transcription, Genetic / Q000187:drug effects*; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
D000911:Antibodies, Monoclonal; D016228:Endothelial Growth Factors; D008222:Lymphokines; D012333:RNA, Messenger; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D000804:Angiotensin II; D013755:Tetradecanoylphorbol Acetate; D019331:NG-Nitroarginine Methyl Ester